Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (3): 275-280.doi: 10.3736/jcim20110307

• Original Clinical Research • Previous Articles     Next Articles

Identification of salivary biomarkers in breast cancer patients with thick white or thick yellow tongue fur using isobaric tags for relative and absolute quantitative proteomics

Mei-qun Cao1,3, Zheng-zhi Wu1,2(), Wei-kang Wu3   

  1. 1. Shenzhen Institute of Integrated Traditional Chinese and Western Medicine, Shenzhen Second People’s Hospital, Shenzhen 518035, Guangdong Province, China;
    2. Shenzhen Institute of Geriatric Medicine, Shenzhen 518020, Guangdong Province, China
    3. Department of Pathophysiology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510080, Guangdong Province, China
  • Received:2010-11-11 Accepted:2010-12-27 Online:2011-03-20 Published:2011-03-15
  • Contact: Wu Zheng-zhi

Objective: To explore the presence of informative protein biomarkers in the salivary proteome of breast cancer patients with thick white or thick yellow tongue fur.
Methods: Salivia samples were collected from 20 breast cancer patients with thick white or yellow tongue fur and 10 healthy controls. The samples were profiled by using isobaric tags for relative and absolute quantitation (iTRAQ) technology coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The analyzed map and data were assessed with Mascott 2.2 and Scaffold software. Ratio of proteins between groups of less than 0.6 or more than 1.5 could confirm that there was difference between groups.
Results: A total of 464 proteins were identified and 125 proteins met strict quantitative criteria. There were 9 proteins associated with breast cancer, expression levels of which were up- or down-regulated more than 1.5 folds compared with healthy people. There were 16 proteins associated with tongue coating, of which 10 proteins expressed in breast cancer patients with thick white fur were lower than in patients with thick yellow fur, and the expressions of the other 6 proteins were increased.
Conclusion: This study demonstrates that iTRAQ combined with LC-MS/MS quantitative proteomics is a powerful tool for biomarker discovery and the identification of proteins associated with breast cancer and tongue coating.

Key words: breast cancer, salivary, isobaric tags for relative and absolute quantitation, proteomics, tongue coating, clinical trial

[1] Jin F . Trends of breast cancer from 1972 to 1992 in Shanghai[J]. Zhongguo Zhong Liu, 1996,5(6):8
金凡 . 上海市区1972年—1992年乳腺癌发病趋势[J]. 中国肿瘤, 1996,5(6):8
[2] Ren XH . Saliva secretion and its influencing factors[J]. Zhongguo Shi Yong Yi Yao, 2009,4(20):252-254
doi: 10.3969/j.issn.1673-7555.2009.20.202
任小红 . 唾液的分泌及影响因素[J]. 中国实用医药, 2009,4(20):252-254
doi: 10.3969/j.issn.1673-7555.2009.20.202
[3] Ross PL, Huang YN, Marchese JN, Williamson B, Parker K, Hattan S, Khainovski N, Pillai S, Dey S, Daniels S, Purkayastha S, Juhasz P, Martin S, Bartlet-Jones M, He F, Jacobson A, Pappin DJ . Multiplexed protein quantitation in Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents[J]. Mol Cell Proteomics, 2004,3(12):1154-1169
doi: 10.1074/mcp.M400129-MCP200
[4] Coldman A, Phillips N, Warren L, Kan L . Breast cancer mortality after screening mammography in British Columbia women[J]. Int J Cancer, 2007,120(5):1076-1080
doi: 10.1002/ijc.22249 pmid: 17149701
[5] Streckfus CF, Mayorga-Wark O, Arreola D, Edwards C, Bigler L, Dubinsky WP . Breast cancer related proteins are present in saliva and are modulated secondary to ductal carcinoma in situ of the breast[J]. Cancer Invest, 2008,26(2):159-167
doi: 10.1080/07357900701783883
[6] De Giovanni N, Fucci N, Chiarotti M, Scarlata S . Cozart Rapiscan system: our experience with saliva tests[J]. J Chromatogr B Analyt Technol Biomed Life Sc, 2002,773(1):1-6
doi: 10.1016/S0378-4347(01)00522-9 pmid: 12015264
[7] Streckfus CF, Bigler LR . Saliva as a diagnostic fluid[J]. Oral Dis, 2002,8(2):69-76
doi: 10.1034/j.1601-0825.2002.1o834.x pmid: 1882670
[8] Ohshiro K, Rosenthal DI, Koomen JM, Streckfus CF, Chambers M, Kobayashi R, El-Naggar AK . Pre-analytic saliva processing affect proteomic results and biomarker screening of head and neck squamous carcinoma[J]. Int J Oncol, 2007,30(3):743-749
[9] Davidovich E, Davidovits M, Peretz B, Shapira J, Aframian DJ . The correlation between dental calculus and disturbed mineral metabolism in paediatric patients with chronic kidney disease[J]. Nephrol Dial Transplant, 2009,24(8):2439-2445
doi: 10.1093/ndt/gfp101
[10] Yan W, Apweiler R, Balgley BM, Boontheung P, Bundy JL, Cargile BJ, Cole S, Fang X, Gonzalez-Begne M, Griffin TJ, Hagen F, Hu S, Wolinsky LE, Lee CS, Malamud D, Melvin JE, Menon R, Mueller M, Qiao R, Rhodus NL, Sevinsky JR, States D, Stephenson JL, Than S, Yates JR, Yu W, Xie H, Xie Y, Omenn GS, Loo JA, Wong DT . Systematic comparison of the human saliva and plasma proteomes[J]. Proteomics Clin Appl, 2009,3(1):116-134
doi: 10.1002/prca.200800140 pmid: 19898684
[11] Wu ZZ, Guo ZQ, Li XH, Zhou XQ . Comprehensive experimental study on the principles of common furs[J]. Zhongguo Zhong Yi Yao Ke Ji, 1996,3(4):5-8
吴正治, 郭振球, 李新华, 周小青 . 舌苔原理的综合实验研究[J]. 中国中医药科技, 1996,3(4):5-8
[12] Wu ZZ, Li M, Zhang YF, Chen MY . Study on relationship between the thickness of tongue fur and the expressions of apoptosis-related genes of the tongue epithelial cells in patients with diseases of the digestive system[J]. J Tradit Chin Med, 2007,27(2):148-152
[13] Li SM, Deng TT . Analysis of salivary amylase activity and xylose excretion rate in myasthenia gravis with spleen-asthenia[J]. Guangzhou Zhong Yi Xue Yuan Xue Bao, 1991,8(4):270-272
李顺民, 邓铁涛 . 重症肌无力脾虚证唾液淀粉酶活性及D-木糖排泄率分析[J]. 广州中医学院学报, 1991,8(4):270-272
[14] Wu ZZ, Wang JG, Zhang XL, Cao MQ . Preliminary study on saliva proteomics of different furs in digestive system diseases. Jinan: IT in Medicine & Education, 2009. ITIME ’09. IEEE International Symposium on. 2009: 56-61.
doi: 10.1109/ITIME.2009.5236461
[15] Li XF, Lin HX, Huang WX, Chen YZ, Jiang H, Lin QC, Wu SS, Chen JZ . A study of apolipoprotein as the index of deficiency and excess syndrome of malignant tumor[J]. Xin Zhong Yi, 2001,33(10):28-29
李晓峰, 林禾禧, 黄伟贤, 陈耀祖, 江红, 林求诚, 吴水生, 陈劲智 . 载脂蛋白作为恶性肿瘤虚实辨证指标的研究[J]. 新中医, 2001,33(10):28-29
[16] Gabay C, Kushner I . Acute-phase proteins and other systemic responses to inflammation[J]. N Engl J Med, 1999,340(6):448-454
doi: 10.1056/NEJM199902113400607 pmid: 9971870
[17] Lecchi C, Avallone G, Giurovich M, Roccabianca P, Ceciliani F . Extra hepatic expression of the acute phase protein alpha 1-acid glycoprotein in normal bovine tissues[J]. Vet J, 2009,180(2):256-258
doi: 10.1016/j.tvjl.2007.12.027
[18] Tang WX, Wang W, Chen ZW, Chen XP . Thioredoxin: a novel targets for tumor therapy[J]. Xian Dai Sheng Wu Yi Xue Jin Zhan, 2009,9(13):2558-2561
唐文心, 王伟, 陈正望, 陈孝平 . 硫氧还蛋白:一个肿瘤治疗的新靶点[J]. 现代生物医学进展, 2009,9(13):2558-2561
[19] Urig S, Becker K . On the potential of thioredoxin reductase inhibitors for cancer therapy[J]. Semin Cancer Biol, 2006,16(6):452-465
doi: 10.1016/j.semcancer.2006.09.004 pmid: 17056271
[20] Baker A, Payne CM, Briehl MM, Powis G . Thioredoxin, a gene found overexpressed in human cancer, inhibits apoptosis in vitro and in vivo[J]. Cancer Res, 1997,57(22):5162-5167
[21] Kos J, Krasovec M, Cimerman N, Nielsen HJ, Christensen IJ, Brünner N . Cysteine proteinase inhibitors stefin A, Stefin B, and cystatin C in sera from patients with colorectal cancer: relation to prognosis[J]. Clin Cancer Res, 2000,6(2):505-511
[22] Hall A, H kansson K, Mason RW, Grubb A, Abrahamson M .Structural basis for the biological specificity of cystatin C. Identification of leucine 9 in the N-terminal binding region as a selectivity-conferring residue in the inhibition of mammalian cysteine peptidases[J]. J Biol Chem, 1995,270(10):5115-5121
doi: 10.1074/jbc.270.10.5115
[23] Parker BS, Ciocca DR, Bidwell BN, Gago FE, Fanelli MA, George J, Slavin JL, Mller A, Steel R, Pouliot N, Eckhardt BL, Henderson MA, Anderson RL .Primary tumour expression of the cysteine cathepsin inhibitor Stefin A inhibits distant metastasis in breast cancer[J]. J Pathol, 2008,214(3):337-346
doi: 10.1002/path.2265
[24] Kuopio T, Kankaanranta A, Jalava P, Kronqvist P, Kotkansalo T, Weber E, Collan Y . Cysteine proteinase inhibitor cystatin A in breast cancer[J]. Cancer Res, 1998,58(3):432-436
doi: 10.1016/S0016-0032(24)90506-0 pmid: 9458085
[1] Pratya Phetkate, Tanawan Kummalue, Prasob-orn Rinthong, Somboon Kietinun, Kusuma Sriyakul. Study of the safety of oral Triphala aqueous extract on healthy volunteers. Journal of Integrative Medicine, 2020, 18(1): 35-40.
[2] Arthur Yin Fan, Hui Ouyang, Xinru Qian, Hui Wei, David Dehui Wang, Deguang He, Haihe Tian, Changzhen Gong, Amy Matecki, Sarah Faggert Alemi. Discussions on real-world acupuncture treatments for chronic low-back pain in older adults. Journal of Integrative Medicine, 2019, 17(2): 71-76.
[3] Carlos R. Oliveira, Daniel G. Spindola, Daniel M. Garcia, Adolfo Erustes, Alexandre Bechara, Caroline Palmeira-dos-Santos, Soraya S. Smaili, Gustavo J.S. Pereira, André Hinsberger, Ezequiel P. Viriato, Maria Cristina Marcucci, Alexandra C.H.F. Sawaya, Samantha L. Tomaz, Elaine G. Rodrigues, Claudia Bincoletto. Medicinal properties of Angelica archangelica root extract: Cytotoxicity in breast cancer cells and its protective effects against in vivo tumor development. Journal of Integrative Medicine, 2019, 17(2): 132-140.
[4] L. Elisabeth Burton, Fares Qeadan, Mark R. Burge. Efficacy of equine-assisted psychotherapy in veterans with posttraumatic stress disorder. Journal of Integrative Medicine, 2019, 17(1): 14-19.
[5] Ubiratan Cardinalli Adler, Ana Carolina Acorinte, Fernando Oliveira Calzavara, Adriano Andréda Silva, Amarilys de Toledo Cesar, Maristela Schiabel Adler, Edson Zangiacomi Martinez, José Carlos Fernandes Galduróz. Double-blind evaluation of homeopathy on cocaine craving: A randomized controlled pilot study. Journal of Integrative Medicine, 2018, 16(3): 178-184.
[6] Shahnaz Karkon Varnosfaderani, Fataneh Hashem-Dabaghian, Gholamreza Amin, Mahbubeh Bozorgi, Ghazaleh Heydarirad, Esmaeil Nazem, Mohsen Nasiri Toosi, Seyed Hamdollah Mosavat. Efficacy and safety of Amla (Phyllanthus emblica L.) in non-erosive reflux disease: A double-blind, randomized, placebo-controlled clinical trial. Journal of Integrative Medicine, 2018, 16(2): 126-131.
[7] Lizhou Liu, Simone Petrich, Blair McLaren, Lyndell Kelly, G. David Baxter. An integrative Tai Chi program for patients with breast cancer undergoing cancer therapy: Study protocol for a randomized controlled feasibility study. Journal of Integrative Medicine, 2018, 16(2): 99-105.
[8] Maryam Sharifi-Ardani, Leili Yekefallah, Saeed Asefzadeh, Marjan Nassiri-Asl. Efficacy of topical chamomile on the incidence of phlebitis due to an amiodarone infusion in coronary care patients: A double-blind, randomized controlled trial. Journal of Integrative Medicine, 2017, 15(5): 373-378.
[9] Lee Suan Chua, Siti Najihah Zukefl i. A comprehensive review of edible bird nests and swiftlet farming. Journal of Integrative Medicine, 2016, 14(6): 415-428.
[10] Gloria Gronowicz, Eric R. Secor Jr., John R. Flynn, Liisa T. Kuhn. Human biofield therapy does not affect tumor size but modulates immune responses in a mouse model for breast cancer. Journal of Integrative Medicine, 2016, 14(5): 389-399.
[11] Susana Seca, Sebastian Kirch, António S.Cabrita, Henry J.Greten. Evaluation of the effect of acupuncture on hand pain, functional deficits and health-related quality of life in patients with rheumatoid arthritis—A study protocol for a multicenter, double-blind, randomized clinical trial. Journal of Integrative Medicine, 2016, 14(3): 219-227.
[12] Arthur Yin Fan. The methodology flaws in Hinman’s acupuncture clinical trial, Part III: Sample size calculation. Journal of Integrative Medicine, 2015, 13(4): 209-211.
[13] Arthur Yin Fan. The methodology flaws in Hinman’s acupuncture clinical trial, Part II: Zelen design and effectiveness dilutions. Journal of Integrative Medicine, 2015, 13(3): 136-139.
[14] Arthur Yin Fan. The methodology flaws in Hinman’s acupuncture clinical trial, Part I: Design and results interpretation. Journal of Integrative Medicine, 2015, 13(2): 65-68.
[15] Vamsi Reddy, Arvind Sridhar, Roberto F. Machado, Jiwang Chen. High sodium causes hypertension: Evidence from clinical trials and animal experiments. Journal of Integrative Medicine, 2015, 13(1): 1-8.
Full text



[1] Jun Cai, Hua Wang, Sheng Zhou, Bin Wu, Hua-rong Song, Zheng-rong Xuan. Effect of Sijunzi Decoction and enteral nutrition on T-cell subsets and nutritional status in patients with gastric cancer after operation: A randomized controlled trial. Journal of Chinese Integrative Medicine, 2008, 6(1): 37-40
[2] SUN Ming-yu, ZUO Jian, DUAN Ji-feng, HAN Jun, FAN Shi-ming, ZHANG Wei, ZHU Li-fang, YAO Ming-hui. Antitumor activities of kushen flavonoids in vivo and in vitro. Journal of Chinese Integrative Medicine, 2008, 6(1): 51-59
[3] Guo-hong Yuan, Xiao-jing Pang, He-chao Ma. Synergic effects of Danggui Buxue Decoction in reducing toxicity of cytoxan in tumor-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 83-88
[4] Bo Wang , Wei Yan , Li-hui Hou, Xiao-ke Wu. Disorder of Tiangui (kidney essence) and reproductive dysfunction in patients with polycystic ovary syndrome. Journal of Chinese Integrative Medicine, 2010, 8(11): 1018-1022
[5] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035
[6] Bing Hu , Hong-mei An , Ke-ping Shen , Qin Du. Senescence-inducing effects of Chinese herbal medicine Tenglong Buzhong Decoction on human colon carcinoma LS-174-T cells and the mechanism. Journal of Chinese Integrative Medicine, 2010, 8(11): 1048-1052
[7] Zhi-mei Wang, Bin Zhang. A study on translation of ellipses in Huangdi Neijing from perspective of hermeneutic theory. Journal of Chinese Integrative Medicine, 2010, 8(11): 1097-1100
[8] Hui-min Liu, Xian-bo Wang, Yu-juan Chang, Li-li Gu. Systematic review and meta-analysis of randomized controlled trials of integrative medicine therapy for treatment of chronic severe hepatitis. Journal of Chinese Integrative Medicine, 2012, 10(11): 1211-1228
[9] Ming-qing Wei , Jin-zhou Tian , Jing Shi , Fu-yun Ma , Ying-chun Miao , Yong-yan Wang. Effects of Chinese medicine for promoting blood circulation and removing blood stasis in treating patients with mild to moderate vascular dementia: a randomized, double-blind and parallel-controlled trial. Journal of Chinese Integrative Medicine, 2012, 10(11): 1240-1246
[10] Nian-hong Wang , Jun-tao Yan , Wu-quan Sun , Yong-shan Hu , Jun Xia , Li-cheng Wei , Jie Jia , Gui-lin Ouyang , Yong He , Yan-ming Guo , Jie Xu . Effects of early application of Tuina treatment on quadriceps surface myoelectricity in patients after total knee arthroplasty: a randomized controlled trial. Journal of Chinese Integrative Medicine, 2012, 10(11): 1247-1253